Development of chromosome sorting techniques using a dual laser chromosome sorter

我们用双激光染色体分选仪分析了人类染色体的结构，用Hoechst 33258（Ho）和色霉素A_3（CA_3）染色的人类染色体依次通过两束激光，一束激光激发CA_3，另一束激光激发Ho，波长分别为457.9nm和351.1-363.8nm。单个染色体的Ho和CA 3含量通过测量当它们通过两个激光束时发射的荧光强度来确定。分选技术已经从用单个激光系统分选10-20个人类染色体片段的能力发展到用双激光系统分选包括15种独特类型的21个片段。不能从分离自人细胞的染色体中以高纯度分选的染色体类型可以从人-含有一条或几条人类染色体的仓鼠杂交细胞系。

We have developed chromosome sorting techniques using a dual laser chromosome sorter to analyze the structure of human chromosomes.Human chromosomes stained with Hoechst 33258 (Ho) and chromomycin A3 (CA3) flowed sequentially through two laser beams, one adjusted to 457.9nm to excite CA3 and the other adjusted to 351.1-363.8nm to excite Ho. The Ho and CA3 contents of individual chromosomes were determined by measuring the intensities of fluorescence that were emitted as they passed two laser beams. These measurements were accumulated to form fluorescence intensity distributions.Sorting techniques have evolved from the ability to sort 10-20 human chromosome fractions with a single laser system to sorting 21 fractions including 15 unique types with a dual laser system.dFurthermore, chromosome types that cannot be sorted at high purity from chromosomes isolated from human cells can be sorted from human-hamster hybrid cell lines containing one or a few human chromosomes.